Geodate delivery vehicles

ABSTRACT

The present invention provides geodate delivery vehicles and methods of manufacture and administration. A vehicle including a lipid monolayer disposed about a hydrophobic domain is disclosed, that can be part of an emulsion or other mixture, or further disposed in a lipid strata. A vehicle including a lipid strata disposed about a hydrophobic domain is also disclosed. The vehicle can be incorporated into a variety of medicinal, food preparations, and personal care products to deliver or stabilize a cargo moiety. Packaged delivery vehicles for later addition of cargo moieties are also contemplated.

RELATED APPLICATIONS

[0001] This application claims the benefit of U.S. ProvisionalApplication No. 60/422,989, filed Nov. 1, 2002, 60/440,284, filed Jan.14, 2003 and 60/507,361, filed Sep. 29, 2003, which applications areincorporated herein by this reference.

BACKGROUND

[0002] Liposomes are widely described in the literature, and theirstructure is well known. Typically, they have an onion-likemultilamellar structure comprising a plurality of lipid bilayers spacedone from another by aqueous material. Another type of liposome is aunilamellar liposome, sometime referred to as a vesicle, which is asingle lipid bilayer disposed about an aqueous material.

[0003] The use of liposomes as carriers or vehicles for drugs is known,and can be achieved by a variety of methods. One method involves castinga film of lipid by evaporation from a solution in an organic solvent,for example chloroform, and then dispersing the film in a suitableaqueous medium. In the case of lipid-soluble biologically activecompounds, that is, those which associate with the lipid layers ratherthan the aqueous phase of the liposomes, the compound can be cast as afilm together with a phospholipid, using a common organic solvent. Adisadvantage of this method is that the amount of active compound thatcan be incorporated into the lipid bilayer is limited. Additionally, thecasting method can not be scaled up to accommodate large batches. In thecase of water-soluble biologically active compounds the compound istypically associated with liposomes by dispersing a cast lipid film withan aqueous solution in which the compound is solubilized. Disadvantagesof this method include the difficulty of incorporating sufficientquantities of the active compound in the vesicles, and instability andshelf life of the dispersion. Another disadvantage of this method is thepresence of trace amounts of solvent used in the creation of thevesicles.

[0004] The loss of the biologically active compound from liposomes intoexternal aqueous medium is another factor which restricts the potentialof these preparations as practical dosage forms. This is particularlythe case for not only low molecular weight, water-soluble compounds, butalso for lipid-soluble compounds, both of which can partition into theexternal aqueous medium until equilibrium is reached. If theconcentration of compound is small, and/or the volume of the externalaqueous medium is large, this loss can represent a significantproportion of the total amount of the biologically active compound inthe liposomes.

SUMMARY OF THE INVENTION

[0005] The present invention provides new delivery vehicles for cargomoieties that are stable and capable of delivering desired amounts ofactive agent. The present invention is based, at least in part, on thediscovery that geodate delivery vehicles can be formed that include alipid monolayer formed about a hydrophobic domain. The hydrophobicdomain can include one or more cargo moieties at concentrationspreviously unattainable by incorporating hydrophobic agents intoliposomal bilayers.

[0006] The present invention also is based, in part, on the discoverythat the delivery vehicles can be locked within a crystal strata ofalternating cation and lipid sheet layers. The encrustation canoptionally be removed prior to administration or administered in anencrusted state. The present invention further provides novel methods ofmanufacture of the, delivery vehicles, including vehicles in emulsionand crystallized form. Encrusted or crystallized vehicles can beconveniently and stably added to further preparations, such as food, andretain their integrity until ingested, retaining the cargo moiety in astable, non-degraded state. Methods of administration and incorporationare also disclosed.

[0007] Thus, in one embodiment, the present invention provides a geodatedelivery vehicle for a cargo moiety which includes a lipid monolayerdisposed about a hydrophobic domain and a lipid strata disposed aboutthe lipid monolayer. In another embodiment, the invention provides ageodate delivery vehicle for a cargo moiety which includes a lipidmonolayer disposed about a hydrophobic domain, wherein the lipidmonolayer includes a phospholipid.

[0008] In some embodiments, the geodate delivery vehicle of the presentinvention is suspended in an aqueous environment. Additionally oralternatively, the geodate delivery vehicle is suspended in an emulsion.In another embodiment, the geodate delivery vehicle is in powder form.

[0009] In some embodiments, the geodate delivery vehicle of includes acargo moiety associated with the geodate delivery vehicle. In oneembodiment, the cargo moiety is associated with the hydrophobic domain.In another embodiment, the hydrophobic domain is a cargo moiety. In athird embodiment, the cargo moiety is associated with the lipidmonolayer or the lipid strata. In a fourth embodiment, the hydrophobicdomain includes a cargo moiety associated with an oil or fat. Inpreferred embodiments, the cargo moiety is a vitamin, a mineral, anutrient, a micronutrient, an amino acid, a toxin, a microbicide, amicrobistat, a co-factor, an enzyme, a polypeptide, a polypeptideaggregate, a polynucleotide, a lipid, a carbohydrate, a nucleotide, astarch, a pigment, a fatty acid, a monounsaturated fatty acid, apolyunsaturated fatty acid, a flavor substance, a flavored essential oilor extract, a hormone, a cytokine, a virus, an organelle, a steroid orother multi-ring structure, a saccharide, a metal, a metabolic poison,an antigen, an imaging agent, a porphyrin, a tetrapyrrolic pigment, or adrug.

[0010] In one aspect of the invention, the lipid includes a negativelycharged phospholipid. Preferably, the lipid includes at least about 50%jnegatively charged lipid, and more preferably, the lipid includes atleast about 75% negatively charged lipid.

[0011] In some embodiments, the geodate delivery vehicle includes anaggregation inhibitor. Preferably, the aggregation inhibitor is caseinor methylcellulose.

[0012] In one aspect, the present invention provides a geodate deliveryvehicle packaged with instructions for incorporating a cargo moiety. Inanother aspect, the invention provides a geodate delivery vehiclepackaged with instructions for adding the vehicle to a food, beverage orpersonal care product.

[0013] In another aspect, the present invention provides a food itemcontaining a geodate delivery vehicle. The food item can be an animalfood item, a human food item, a nutrient bar, a snack food, a beverage,a domesticated animal food, a fish food, a poultry feed, a pet food, adog food or a cat food.

[0014] In yet another aspect, the present invention provides a personalcare product containing a geodate delivery vehicle. The personal careproduct can be a hair care product or a skin care product.

[0015] In other embodiments, the present invention provides apharmaceutical composition including a geodate delivery vehicle and apharmaceutically acceptable carrier.

[0016] In another aspect, the present invention provides a method oftreating a subject that can benefit from the administration of a cargomoiety, comprising the step of administering a geodate delivery vehiclecomprising a cargo moiety to a subject. The route of administration canbe mucosal, systemic, oral, intranasal, intraocular, intrarectal,intravaginal, intrapulmonary, intravenous, intramuscular, subcutaneous,transdermal or intradermal. In some embodiments, the cargo moiety isadministered to treat inflammation, pain, infection, fungal infection,bacterial infection, viral infection, parasitic disorders, an immunedisorder, genetic disorders, degenerative disorders, cancer, diabetes,insomnia, proliferative disorders, obesity, depression, hair loss,impotence, hypertension, hypotension, dementia, senile dementia, ormalnutrition. In other embodiments, the subject can benefit fromadministration of a nutrient and the cargo moiety is a nutrient.

[0017] In another embodiment, the present invention provides a method ofmanufacturing a geodate delivery vehicle for a cargo moiety by mixing alipid, an aqueous solution and a hydrophobic material, such that a lipidmonolayer is disposed about a hydrophobic domain. In another embodiment,the geodate delivery vehicle can additionally include a cargo moiety. Inyet another embodiment, a lipid strata can be formed about the lipidmonolayer by adding a multivalent cation. Preferably, the multivalentcation includes calcium.

[0018] In some embodiments, the geodate delivery vehicle is dried toform a powder. In other embodiments, the geodate delivery vehicle isassociated with a pharmaceutically acceptable carrier. In still otherembodiments, the geodate delivery vehicle is added to a food item or apersonal care product.

[0019] The present invention also provides a method of forming a geodatedelivery vehicle for a cargo moiety by mixing a lipid which includes aphospholipid with a hydrophobic material such that a geodate deliveryvehicle is formed.

BRIEF DESCRIPTION OF THE DRAWINGS

[0020]FIG. 1 illustrates an exemplary method of manufacturing a geodatedelivery vehicle in accordance with the present invention.

[0021] FIGS. 2A-D are four images of dioleoyl phosphatidylserine (DOPS)and olive oil interacting an aqueous buffer.

[0022] FIGS. 3A-B are two images of exemplary geodate delivery vehiclesthat include a DOPS monolayer disposed about Amphotericin B in oliveoil.

[0023]FIG. 4 is an image of a DOPS geodate delivery vehicle thatincludes a DOPS monolayer about fluorescent Amphotericin B interactingwith olive oil.

[0024]FIGS. 5 and 6 are images of a lipid strata encrusting olive oil,where the lipid includes Rhodamine-labeled DOPS.)

[0025]FIGS. 7 and 8 are images of a geodate delivery vehicle including afluorescent DOPS monolayer disposed about olive oil, after release froma lipid strata.

[0026]FIG. 9 illustrates another exemplary method of manufacturing ageodate delivery vehicle in accordance with the present invention.

[0027]FIGS. 10A and B are two images of a stable beta-carotene/oil/lipidemulsion in aqueous media.

[0028]FIG. 11 is an image of a stable emulsion of beta-carotene in soyoil and lipid dispersed in an aqueous environment.

[0029]FIGS. 12A and B are two images of beta carotene-oil geodes made inaccordance with the present invention.

[0030] FIGS. 13A-D are images of beta carotene geodes prepared inaccordance with the present invention. FIGS. 13A and 13B depict geodesin suspension and FIGS. 13C and 13D depict the same formulationssubsequent to the addition of EDTA.

[0031] FIGS. 14A-D are images of beta-carotene geodes extracted fromsuspension in commercial drying apparatus. FIGS. 14A and 14D are imagesof geodes extracted by spray drying, and FIGS. 14B and 14C are images ofgeodes extracted by fluid bed drying.

[0032]FIG. 15 is a graph showing the stability of beta-carotene geodesin suspension, after spray drying, and after fluid bed drying.

[0033]FIG. 16 is two graphs depicting the stability of variousbeta-carotene geode formulations over a 20 day period.

[0034]FIG. 17 is a graph showing the stability of beta-carotene geodesin suspension and after spray drying.

[0035]FIG. 18 is an image of a muffin containing beta-carotene geodes.

[0036]FIG. 19 is an image of beta-carotene geodes applied topically tothe palm.

DETAILED DESCRIPTION OF THE INVENTION

[0037] The invention is based, in part, on the discovery that a lipidmonolayer will form about a hydrophobic domain, and that this structurecan be employed to deliver a variety of cargo moieties.

[0038] One advantage of the present invention is that cargo moieties canbe incorporated into the geodate delivery vehicle at highconcentrations. Another advantage of the present invention is theability to incorporate multiple cargo moieties into one geodate deliveryvehicle. Incorporation into a geodate delivery vehicle is alsoadvantageous because it provides the cargo moiety with protection fromboth the environment, e.g., water and oxygen, and also the stomach.Additionally, the geodate delivery vehicle protects stomach from thecargo moiety. The present invention is advantageous because theformulation of geodate delivery vehicles involves no solvent. Thepresent invention is also advantageous because the resultant geodatedelivery vehicles are highly stable, e.g., they can withstand extremetemperature and pressure. Another advantage of the present invention isthe ability of the geodate delivery vehicle to mask the taste and/orodor of cargo moieties.

[0039] In order to more clearly and concisely describe the subjectmatter of the claims, the following definitions are intended to provideguidance as to the meaning of specific terms used in the followingwritten description, examples and appended claims.

[0040] The term “geodate delivery vehicle” refers to a delivery vehiclefor a cargo moiety. Geodate delivery vehicles generally include a lipidmonolayer disposed about a hydrophobic domain. A “hydrophobic domain” isa composition that is sufficiently hydrophobic in nature to allowformation of a lipid monolayer about its periphery. A hydrophobic domaincan itself be one or more cargo moieties, or it can include ahydrophobic composition, such as oil or fat, associated with the cargomoiety, which can be, e.g., a hydrophobic or amphiphilic agent.

[0041] The term “lipid monolayer” generally refers to a lipid-containinglayer one molecule thick (as contrasted with lipid bilayers that are twomolecules thick). A lipid monolayer can contain further elements, suchas cholesterol, steroids, or proteins. In contrast, “liposomes” refer tovesicles defined by lipid bilayers (two molecules thick) in aunilamellar or multilamellar structure.

[0042] In one aspect of the invention, the lipid monolayer includesand/or is composed primarily of negatively charged lipids. When a lipidstrata is formed, the multivalent cation forms a cationic bridge betweenthe negatively-charged lipid in the monolayer and the negatively chargedlipid in the liposomes. In another embodiment, the lipid monolayer iscomposed primarily of positively charged lipids. In this case, the headgroups interact with negatively charged lipid in the strata. In yetanother embodiment, the lipid monolayer is composed primarily of neutrallipids. The coated hydrophobic domain, in this embodiment is trappedwithin the lipid strata, but does not ionically interact with thestrata.

[0043] The term “lipid strata” refers to a structure of alternatingcationic and lipid sheet-like layers. A lipid strata can be formed byintroducing a cation to an emulsion containing liposomes. The lipidstrata not only locks the hydrophobic domain within the geodate lipidmonolayer, but can itself be associated with a cargo moiety (e.g., ahydrophilic agent disposed within the lipid strata). In one embodiment,the lipid strata entraps a hydrophobic domain. In another, the lipidstrata entraps a hydrophobic domain disposed within a lipid monolayer.

[0044] The term “cargo moiety” refers to any compound having a propertyof biological interest. The agent may be, e.g., organic or inorganic, amonomer or a polymer, endogenous to a host organism or not, naturallyoccurring or synthesized in vitro and the like. Thus, examples include,vitamins, minerals, nutrients, micronutrients, amino acids, toxins,microbicides, microbistats, co-factors, enzymes, polypeptides,polypeptide aggregates, polynucleotides, lipids, carbohydrates,nucleotides, starches, pigments, fatty acids, monounsaturated fattyacids, polyunsaturated fatty acids, flavorings, essential oils orextracts, hormones, cytokines, viruses, organelles, steroids and othermulti-ring structures, saccharides, metals, medicaments, proteins,marker compounds, imaging agents, antigens, porphyrins, tetrapyrrolicpigments, metabolic poisons, drugs and the like

[0045] The methods of this invention are particularly useful in the caseof hydrophobic cargo moieties and agents that can be associated withand/or can be incorporated into a hydrophobic phase, e.g., by binding toor admixing with a hydrophobic vehicle. Cargo moieties can also beincorporated in a lipid strata of the present invention. Thus,combination therapies can be employed by delivering one or more activeagents (e.g., hydrophobic and amphiphilic agents) associated with thehydrophobic domain, and one or more active agents (e.g., hydrophilicagents) associated with the lipid strata.

[0046] In one embodiment, the invention provides a geodate deliveryvehicle for a cargo moiety, which includes a lipid monolayer disposed ahydrophobic domain, and a lipid strata disposed about the lipidmonolayer. In another embodiment, the invention provides a geodatedelivery vehicle for a cargo moiety including a lipid monolayer disposeda hydrophobic domain, and wherein the lipid monolayer includes at leastone phospholipid.

[0047] In one embodiment, the geodate delivery vehicle can be suspendedin an aqueous environment, e.g., an emulsion. In alternativeembodiments, the geodate delivery vehicle is in powder form.

[0048] The hydrophobic domain is a hydrophobic composition that can be acarrier for one or more cargo moieties, or the cargo moiety or agentsitself. That is, the hydrophobic domain can be a hydrophobic carrier(e.g., olive oil or soy oil) associated with a cargo moiety (e.g., anantifungal agent such as amphotericin, a marker compound such asrhodamine, and/or nutrients such as beta carotene and alpha tocopherol).Alternatively, the hydrophobic domain can be the cargo moiety itself,e.g., a nutrient such as omega 3 fatty acid or a hydrophobic drug.Alternatively the hydrophobic domain can be one or more cargo moietiesthat act as a carrier for further cargo moieties.

[0049] In one embodiment, the hydrophobic domain is present in a rangeof between about 1% and 99%, preferably between about 1% and about 75%,more preferably between about 10% and about 30% by weight of the finalcomposition of the geode.

[0050] The terms “encrusted,” “crystallized,” and “crystalline”generally refer to a solid or semi-solid lipid strata formed about oneor more hydrophobic domains.

[0051] As used herein, the term “food” refers to any object or objectssuitable for consumption by a human or non-human animal.

[0052] The term “delivery,” as used herein, refers to any means ofbringing or transporting a cargo moiety to a host, a food item, aformulation, a pharmaceutical composition, or any other system, whereinthe cargo moiety maintains at least a portion of the activity it hadwhen first formulated in the geodate structure. Thus, e.g., in abeta-carotene geode, the beta-carotene retains some activity within thegeode until it is used.

[0053] The delivery vehicles of the present invention are directed tohydrophobic domains encapsulated or entrapped in a stable vehicle. Inone aspect, the invention features a lipid monolayer disposed about ahydrophobic domain, which can be part of a stable emulsion and/orentrapped in lipid strata. In another aspect the geodate deliveryvehicle features a lipid strata disposed about a hydrophobic domain, anda lipid monolayer is optional.

[0054] The hydrophobic domain can itself be one or more cargo moieties,or it can include a hydrophobic composition (e.g., oil or fat)associated with the cargo moiety, which can be, e.g., a hydrophobic oramphiphilic agent. If the agent is amphiphilic, it can associate withboth the hydrophobic domain and the lipid. Further cargo moieties canalso be delivered by associating them with a lipid strata, thuscombination therapies can be effected. The cargo moiety can beassociated with the hydrophobic domain, the lipid monolayer and/or thelipid strata.

[0055] The cargo moiety can be a diagnostic agent, such as an imagingagent. Imaging agents include nuclear agents and porphyrins. Porphyrinsinclude tetrapyrrolic agents or pigments. One such tetrapyrrolic agentis Zinc Tetra-Phenyl Porphyrin (ZnTPP), which is a hydrophobic,fluorescent molecule that has high absorption in the visible spectrum(dark purple).

[0056] The cargo moiety may be a polynucleotide that is expressed toyield a biologically active polypeptide or polynucleotide. Thus, thepolypeptide may serve as an immunogen or, e.g., have enzymatic activity.The polynucleotide may have catalytic activity, for example, be aribosome, or may serve as an inhibitor of transcription or translation,e.g., a small interfering RNA (siRNA) or an antisense molecule. Thepolynucleotide can be an antisense molecule including modified antisensemolecule, such as an morpholino antisense molecule. The polynucleotidecan be modified, e.g., it can be synthesized to have a morpholinobackbone. If expressed, the polynucleotide preferably includes thenecessary regulatory elements, such as a promoter, as known in the art.A specific example of a polypeptide is insulin. The cargo moiety can bean organic molecule that is hydrophobic in aqueous media. The cargomoiety can be a water-soluble polyvalent cationic molecule.

[0057] The cargo moiety can be a drug, such as, a protein, a smallpeptide, a bioactive polynucleotide, an antibiotic, an antiviral, ananesthetic, an anti-infectious, an antifungal, an anticancer, animmunosuppressant, a steroidal anti-inflammatory, a non-steroidalanti-inflammatory, an antioxidant, an antidepressant which can besynthetic or naturally derived, a substance which supports or enhancesmental function or inhibits mental deterioration, an anticonvulsant, anHIV protease inhibitor, a non-nucleophilic reverse transcriptaseinhibitor, a cytokine, a tranquilizer or a vasodilatory agent. The drugcan also be any over the counter (non-prescription) medication. Examplesinclude Amphotericin B, acyclovir, adriamycin, carbamazepine,ivermectin, melphalen, nifedipine, indomethacin, curcumin, aspirin,ibuprofen, naproxen, acetaminophen, rofecoxib, diclofenac, ketoprofin,meloxicam, nabumetone, estrogens, testosterones, steroids, phenyloin,ergotamines, cannabinoids, rapamycin, propanadid, propofol, alphadione,echinomycin, miconazole nitrate, teniposide, hexamethylmelamine, taxol,taxotere, 18-hydroxydeoxycorticosterone, prednisolone, dexamethazone,cortisone, hydrocortisone, piroxicam, diazepam, verapamil, vancomycin,tobramycin, nystatin, rifampin, geldanamycin, tyrphostin, glucansynthesis inhibitors, vitamin A acid, mesalamine, risedronate,nitrofurantoin, dantrolene, etidronate, caspofungin, nicotine,amitriptyline, clomipramine, citalopram, dothepin, doxepin, fluoxetine,imipramine, lofepramine, mirtazapine, nortriptyline, paroxetine,reboxitine, sertraline, trazodone, venlafaxine, dopamine, St. John'swort, phosphatidylserine, phosphatidic acid, amastatin, antipain,bestatin, benzamidine, chymostatin, 3,4-dichloroisocoumarin,elastatinal, leupeptin, pepstatin, 1,10-phenanthroline, phosphoramidon,ethosuximide, ethotoin, felbamate, fosphenyloin, lamotrigine,levitiracetam, mephenyloin, methsuximide, oxcarbazepine, phenobarbital,phensuximide, primidone, topirimate, trimethadione, zonisamide,saquinavir, ritonavir, indinavir, nelfinavir, or amprenavir.

[0058] The drug can be a polypeptide such as cyclosporin, angiotensin I,II and III, enkephalins and their analogs, ACTH, anti-inflammatorypeptides I, II, III, bradykinin, calcitonin, b-endorphin, dinorphin,leucokinin, leutinizing hormone releasing hormone (LHRH), insulin,neurokinins, somatostatin, substance P, thyroid releasing hormone (TRH)and vasopressin.

[0059] The drug can be an antigen, but is not limited to a proteinantigen. The antigen can also be a carbohydrate or DNA. Examples ofantigenic proteins include membrane proteins, carbohydrates, envelopeglycoproteins from viruses, animal cell proteins, plant cell proteins,bacterial proteins, and parasitic proteins.

[0060] The antigen can be extracted from the source particle, cell,tissue, or organism by known methods. Biological activity of the antigenneed not be maintained. However, in some instances (e.g., where aprotein has membrane fusion or ligand binding activity or a complexconformation which is recognized by the immune system), it is desirableto maintain the biological activity. In these instances, an extractionbuffer containing a detergent which does not destroy the biologicalactivity of the membrane protein is employed. Suitable detergentsinclude ionic detergents such as cholate salts, deoxycholate salts andthe like or heterogeneous polyoxyethylene detergents such as Tween, BRIGor Triton.

[0061] The cargo moiety can be a nutrient including, but not limited to,lycopene, micronutrients such as phytochemicals or zoochemicals,vitamins, minerals, fatty acids, amino acids, fish oils, fish oilextracts, and saccharides, vitamins, herbal products, essential oils orminerals. Specific examples include Vitamins A, B, B1, B2, B3, B12, B6,B-complex, C, D, E, and K, vitamin precursors, caroteniods, andbeta-carotene, resveratrol, biotin, choline, inositol, ginko, lutein,zeaxanthine, quercetin, silibinin, perillyl alcohol, genistein,sulfurophane, omega-3 and omega-6 fatty acids, herbs, spices, and iron.Minerals include, but are not limited to boron, chromium, colloidalminerals, colloidal silver, copper, manganese, potassium, selenium,vanadium, vanadyl sulfate, calcium, magnesium, barium, iron and zinc.

[0062] As used herein, “micronutrient” is a nutrient that the body mustobtain from outside sources. Generally micronutrients are essential tothe body in small amounts.

[0063] The cargo moiety can be a saccharide or sweetener, e.g.,saccharine, isomalt, maltodextrine, aspartame, glucose, maltose,dextrose, fructose and sucrose. Flavor agents include oils, essentialoils, or extracts, including but not limited to oils and extracts ofcinnamon, vanilla, almond, peppermint, spearmint, chamomile, geranium,ginger, grapefruit, hyssop, jasmine, lavender, lemon, lemongrass,marjoram, lime, nutmeg, orange, rosemary, sage, rose, thyme, anise,basil, black pepper, tea or tea extracts, an herb, a citrus, a spice ora seed.

[0064] In one embodiment, the cargo moiety is present in a range ofbetween approximately 1% and 99% of the final composition. In oneembodiment, the cargo moiety is present in a range between about 1% andabout 30% by weight of the final composition of the geode. In anotherembodiment, a second cargo moiety is additionally incorporated into thegeode structure, in a range of between about 0.1% and about 90% byweight of the final composition of the geode. In one embodiment, thesecond cargo moiety is present in a range of between about 1% and about10%, more preferably between about 1% and about 5%.

[0065] In one embodiment, the cargo moiety is incorporated into thehydrophobic domain in a range of between about 0.1% and about 99% of thehydrophobic domain. In a preferred embodiment, the range is betweenabout 0.1% and about 50%. More preferably, the ratio is between about 1%and about 25%. In another embodiment, a second cargo moiety is alsoincorporated into the hydrophobic domain in a range of between about 1%and about 90%.

[0066] In another embodiment the cargo moiety is the hydrophobic domain.In one embodiment, the hydrophobic domain is present in a range ofbetween about 1% and about 99% of the total composition. In a preferredembodiment, the hydrophobic domain is present in a range of about 1% andabout 50% of the total composition. More preferably, the hydrophobicdomain is present in a range of about 5% to about 35%.

[0067] Lipids suitable for use in forming the lipid monolayer (and theliposomes discussed below) include, but are not limited to,phospholipids such as soy lecithin, partially refined lecithin,hydrogenated phospholipids, lysophosphate, phopshpatidylcholine,phosphatidylethanolamine, phosphatidylserine (PS), phosphatidylinositol,cardiolipin, sphingolipids, gangliosides, cerebrosides, ceramides, soyphospholipids, other ester analogues of phopshpatidylcholine, syntheticphospholipids, phosphatidylethanolamine derivatives, and phospholipidswith partially or fully fluorinated fatty acid chains. Preferably, thelipid is a negatively charged phospholipid such as phosphatidylserine.Preferred phosphatidylserines include soy PS and dioleoyl PS (DOPS). Thelipid can also include fluorescent phospholipid.

[0068] Further, synthetic phospholipids containing either alteredaliphatic portions, such as hydroxyl groups, branched carbon chains,cyclo derivatives, aromatic derivatives, ethers, amides, polyunsaturatedderivatives, halogenated derivatives, or altered hydrophilic portionscontaining carbohydrate, glycol, phosphate, phosphonate, quaternaryamine, sulfate, sulfonate, carboxy, amine, sulfhydryl, imidazole groupsand combinations of such groups, can be either substituted or intermixedwith the phospholipids, and others known to those skilled in the art

[0069] The lipid employed in the present invention preferably includesone or more negatively charged lipids. As used herein, the term“negatively charged lipid” includes lipids having a head group bearing aformal negative charge in aqueous solution at an acidic or physiologicalpH, and also includes lipids having a zwitterionic head group.

[0070] In one embodiment, the lipid is a mixture of lipids, comprisingat least 50% negatively charged lipid. In another embodiment, the lipidincludes at least 75% negatively charged lipid. In other embodiments,the lipid includes at least 85%, 90%, 95% or even 99% negatively chargedlipid. All ranges and values between 40% and 100% negatively chargedlipid are meant to be encompassed herein.

[0071] In a preferred embodiment, the lipid monolayer formed about thehydrophobic domain is a predominantly negatively charged lipidmonolayer. In a preferred embodiment, lipid strata can be formed by theaddition of a cation to the emulsion.

[0072] In another embodiment, the lipid monolayer formed about thehydrophobic domain is a predominantly positively charged lipidmonolayer. In another embodiment, lipid strata may be formed by theaddition of an anion to the emulsion.

[0073] If the delivery vehicle is suspended in a stable emulsion, thesolution can contain liposomes or other lipid structures to furtherstabilize the emulsion, e.g., to reduce or eliminate aggregation orcoalescence within the emulsion. The solution can also includeadditional additives to prevent aggregation, to aid in the associationof cargo moieties with the hydrophobic domains, and/or to prevent theactive agent from migrating out of the delivery vehicles of the presentinvention. If lipid strata is formed about the hydrophobic domains in anemulsion, the vehicles can be utilized in an emulsion or extracted forutility in a solid or semi-solid form such as a paste or a powder.

[0074] The lipid monolayer is advantageous because large and/or chargedmolecules have difficulty passing through it. Thus cargo moieties areinhibited or prevented from exiting the domain through the monolayer.The lipid strata is advantageous because it typically is impassable bycargo moieties which are immuobilized within it or trapped by it in thehydrophobic domain. Another advantage to both the monolayer and thelipid strata is that the cargo moiety is protected from the environmentand the environment is protected by the cargo moiety. Both the emulsionsand the lipid strata are stable, thus enabling not only convenientstorage and delivery of the agents, but a convenient means ofincorporating the same into compositions, such as food or pharmaceuticalcompositions.

[0075] A lipid strata includes liposomes and cation, and can be formedfrom liposomes by exposure to cation. The cation and the liposomes alignto form a stacked or rolled structure that captures and retains orencrusts one or more hydrophobic domains. The cation preferably is amultivalent cation. The cation can be a divalent cation, such as Ca⁺⁺,Zn⁺⁺, Ba⁺⁺, and Mg⁺⁺. The cation can also be a multivalent cargo moiety.

[0076] The hydrophobic domains, with or without a lipid monolayerdispersed about said hydrophobic domains, can be released from the lipidstrata when desired upon exposure of the crystalline lipid structure toa chelating agent such as EDTA, ascorbic acid and/or citric acid. Thechelating agent serves to disrupt the crystalline structure providing ade-encrusted lipid monolayer encapsulating a hydrophobic globule. Thechelating agent can be added to a dry powder and stored, so that uponaddition of water, the chelating agent acts on the encrustation torelease the encapsulated domain.

[0077] Cargo moieties can be delivered at different rates, depending,e.g., on whether the vehicle is in a lipid strata and/or an emulsion.The choice of cation, lipid and hydrophobic domain makeup can alsoaffect delivery rates and times. Thus, the rate of release of the cargomoieties contained therewith varies and can even be staggered, e.g., ifthe lipid strata dissolves first in vivo, delivering a first agent,followed by the delivery of a second agent associated with thehydrophobic domain. Accordingly, by controlling the ingredients and thestructure of the vehicles described herein, vehicles which will releasethe cargo moiety in desired amounts over a protracted period of time areobtainable.

[0078] Accordingly, the compositions of the invention may include one ormore cargo moieties present in or associated with the hydrophobicdomain, the lipid monolayer, the lipid strata, a stable emulsion (e.g.,in liposomes or aqueous media), or any combination thereof. In addition,several layers of precipitate can be formed about or encrusted about thegeodate delivery vehicles, with one or more cargo moieties associatedtherewith. Accordingly, the invention may be employed for combinationdrug therapy and/or consecutive or simultaneous release profiles, e.g.,pulsed or extended release. For example, a stomach protecting medicationcan be formulated in the lipid strata for initial release, and one ormore non-steroidal anti-inflammatory drugs can be formulated in thehydrophobic domain for release after the stomach protecting medicationis released.

[0079] The amount of cargo moiety incorporated into the vehicles of thepresent invention can vary. Because of the advantageous properties ofthe vehicles, e.g., the stability of the agent trapped in the vehicle,lesser amounts of the agent can be used to achieve the same end resultas compared to using known delivery means, e.g., direct addition of theagent to food.

[0080] In one embodiment, the geodate delivery vehicles of the presentinvention are small, e.g., in the micrometer or nanometer range. Suchgeodate delivery vehicles are particularly advantageous, e.g., becausethe small size increases the oral availability. In addition, small sizesare preferred and sometimes necessary for intravenous administration.The geodate delivery vehicles of the invention can be micronized ordisaggregated by introducing an aggregation inhibitor (e.g., casein).Preferably, however, the geodate delivery vehicles are formed in thedesired size range and/or the suspension can be micronized prior toaddition of cation. In such embodiments, an aggregation inhibitor can beemployed to form geodate delivery vehicles in a desired size range.

[0081] In a preferred embodiment the geodate delivery vehicles of thepresent invention further comprise and aggregation inhibitor. In oneembodiment, an aggregation inhibitor is employed to obtain geodatedelivery vehicles of a desired size. The term “aggregation inhibitor,”as used herein, refers to an agent that inhibits aggregation of ageodate delivery vehicle with or without a lipid strata and with orwithout an emulsion. The aggregation inhibitor typically is present atleast on the surface of the geodate delivery vehicle, and may only bepresent on the surface of the geodate delivery vehicle (e.g., when theaggregation inhibitor is introduced after precipitation). Aggregationinhibitors can be added before, after, or during geodate deliveryvehicle formation. Aggregation inhibitors work in part by modifying thesurface characteristics of the geodate delivery vehicle such thataggregation is inhibited. Aggregation can be inhibited, for example, bysteric bulk and/or a change in the nature of the geodate deliveryvehicle structure, e.g., a change in the surface hydrophobicity and/orsurface charge. The aggregation inhibitor can be added at any point inthe manufacture (e.g., to pre-empt aggregation), and/or aftermanufacture (e.g., to stabilize the precipitate size and/or disaggregateprecipitates).

[0082] In a preferred embodiment, the precipitates of the presentinvention include one or more aggregation inhibitors. The aggregationinhibitor can be added prior to, during, and/or after precipitation. Thetype and/or amount of aggregation inhibitor can be adjusted to obtain adesired precipitate size and/or distribution. Additionally oralternatively, aggregation inhibitor(s) can be used to stabilizeprecipitate size and/or size distribution such that aggregation ofprecipitates is minimized or eliminated.

[0083] Suitable aggregation inhibitors that can be employed inaccordance with the present invention, include but are not limited to atleast one of the following: casein, κ-casein, milk, methylcellulose,ethylcellulose, propylcellulose, hydroxycellulose, hydroxymethylcellulose, hydroxyethyl cellulose, hydroxypropyl cellulose,hydroxypropylmethyl cellulose, polyvinyl pyrrolidone, carboxymethylcellulose, carboxyethyl cellulose, pullulan, polyvinyl alcohol, sodiumalginate, polyethylene glycol, polyethylene oxide, xanthan gum,tragacanth gum, guar gum, acacia gum, arabic gum, polyacrylic acid,methylmethacrylate copolymer, carboxyvinyl polymer, amylose, highamylose starch, hydroxypropylated high amylose starch, dextrin, pectin,chitin, chitosan, levan, elsinan, collagen, gelatin, zein, gluten,carrageenan, carnauba wax, shellac, latex polymers, milk proteinisolate, soy protein isolate, whey protein isolate and mixtures thereof.

[0084] A preferred aggregation inhibitor is casein. Casein is a highlyphosphorylated, calcium binding protein. Without wishing to be bound toany particular theory, it is believed that calcium mediates aninteraction between negatively charged lipid (e.g., PS) and casein,thereby changing the surface properties of precipitates such thataggregation is inhibited. Another preferred aggregation inhibitor ismilk and other milk products such as Half and Half, cream, etc. Anotherpreferred aggregation inhibitor is methylcellulose.

[0085] More than one aggregation inhibitor may be employed in thecompositions of the invention. For example, both milk andmethylcellulose may be used as an aggregation inhibitor.

[0086] In one embodiment, the precipitate compositions of the inventioninclude an aggregation inhibitor to lipid ratio of between about 0.5:1to about 4:1 by weight. Preferably, the aggregation inhibitor to lipidratio is about 1:1. A person of ordinary skill in the art will readilybe able to determine the amount of aggregation inhibitor needed to formprecipitates of the desired size with no more than routineexperimentation.

[0087] Pharmaceutical formulations incorporating the delivery vehiclesof the present invention can be of solid form including tablets,capsules, pills, bulk or unit dose powders and granules or of liquidform including solutions, fluid emulsions, fluid suspensions, semisolidsand the like. This is particularly true using vehicles including a lipidstrata, as the crystalline structure protects the agent from itsenvironment and vice versa. In addition to the active ingredient, theformulation would comprise suitable art-recognized diluents, carriers,fillers, binders, emulsifiers, surfactants, water-soluble vehicles,buffers, solubilizers and preservatives.

[0088] Pharmaceutical formulations incorporating the delivery vehiclesof the present invention can be of liquid or semi-liquid form includingfood products, such as therapy or nutrient drinks, yogurt, milk, saladdressing, moist animal food, and the like. The stable emulsions of thepresent invention can be directly added to such formulations.

[0089] An advantage of the vehicles of the present invention is thestability and safety of the composition, particularly when soy-basedlipids are employed. Thus, the geodate delivery vehicles can beadministered orally or by instillation without concern, as well as bythe more traditional routes, such as topical, subcutaneous, intradermal,intramuscular and the like. Direct application to mucosal surfaces is anattractive delivery means made possible with the delivery vehicles.

[0090] The skilled artisan can determine the most efficacious andtherapeutic means for effecting treatment practicing the instantinvention. Reference can also be made to any of numerous authorities andreferences including, for example, “Goodman & Gilman's, ThePharmaceutical Basis for Therapeutics”, (6th Ed., Goodman, et al., eds.,MacMillan Publ. Co., New York, 1980).

[0091] The geodate delivery vehicles of the instant invention also serveas excellent means for delivering fragile cargo moieties to a host. Suchcargo moieties include nutrients, vitamins such as vitamins A, D, E orK, co-factors, enzymes, fatty acids such as polyunsaturated forms,minerals including divalent cations such as calcium, magnesium, zinc,iron or barium, flavors and the like. Because the cargo moiety iscontained within the vehicle, in a non-aqueous environment, the agentessentially is stabilized and preserved. Hydrophobic molecules can bemade part of the geodate delivery structure, with little difficulty asthe lipid monolayer of the present invention will form about ahydrophobic domain.

[0092] The geodate delivery vehicles can be particularly advantageousfor delivering agents to food and drinks to be consumed by humans orother animals. For example, dog and cat food can include the vehicles ofthe present invention to stably deliver vitamins, flavoring agents,minerals or other nutrients, as well as medications, e.g., allergymedications. Similarly, the geodate delivery vehicles of the presentinvention can be added to pet or domestic animal feed, such as fish foodand food for fowl, cattle, and horses. The vehicles can be added at anystep of the preparation. For example, the vehicles can be added at anypoint in the methods described in WO 02/44026, incorporated herein bythis reference. Similarly, the compositions and methods of the inventioncan be employed in food or drink to be consumed by humans, e.g., in anutrient bar or drink, cereals, breads, and snack food. Accordingly, thepreparations of the invention allow for the production of stable,convenient preparations of micronutrients in processed foods, such asfast foods. Typically, potentially beneficial micronutrients, e.g.,omega fatty acids and antioxidants, can be destroyed during foodmanufacture and storage. The delivery vehicles of the invention protectmicronutrients and other cargo moieties, thus increasing the nutritionaland/or medicinal value of the food.

[0093] Because of their increased stability, the compositions andmethods of the present invention are particularly useful in foods thatare baked or cooked, such as cakes, muffins, pasta noodles, soups,cereals, chips, candy and cookies. In a preferred embodiment, thecompositions are used in candy, such as candy bars, e.g., chocolatebars. For example, omega fatty acid-geodes can be incorporated into achocolate bar.

[0094] The geodate delivery vehicles can be added to food items, e.g.,fast food products, in the crystallized or emulsion form at any stage ofthe manufacturing process. The food item can be an animal food item, ahuman food item, a nutrient bar, a snack food, a beverage, adomesticated animal food, a fish food, a poultry feed, a pet food, a dogfood or a cat food

[0095] They preferably are added at a stage where the integrity of thedelivery vehicle is maintained until ingestion, or final preparation ofthe food product by the consumer. Another alternative, however, can beto use the vehicles to maintain the stability of the agent untilincorporation into the product, so activity can be maintained duringstorage and shipping. Yet another alternative is to deliver the vehiclesthemselves to consumers or professionals, for direct addition to foodproducts, e.g., medicament, nutrient crystals, additives, supplements,or emulsions, such that the user can vary the concentration as desired.

[0096] The vehicles can also be added to a carrier for use as a topicaltreatment on the skin. Suitable carriers would remain on the skin for anextended period of time, and be resistant to perspiration or immersionin water. Thus, for example, the vehicles may be added to topicalapplications of medicaments, moisturizers, deodorants, balms,fragrances, sunscreens, and the like.

[0097] Additional examples of formulations that can include the geodatedelivery vehicles of the invention include, but are not limited to, haircare products, skin care products, personal care products, personalcleansing products, lotions, fragrances, sprays, perfumes, cosmetics,toothpastes, tooth whiteners, cleaners, bar soap, liquid soap, bodywash, baby wash, makeup, hair color, shampoos, conditioners, stylingproducts, balms, creams, solutions, gels and solids. Thus, for example,shampoos, conditioners and the like may contain geodate deliveryvehicles loaded with vitamins, moisturizers, perfumes, medications, etc.

[0098] The vehicles can also be added to cleansers which do not havedirect contact with the skin. These formulations would be advantageousfor, i.e., the incorporation of perfumes, moisturizers or other suchcargo moieties into fabric or for the introduction of an antibacterialagent to dishes. Examples include, but are not limited to, laundrydetergent, pre-treating formulations, dryer sheets, fabric softener, anddishwashing detergent.

[0099] Geodate delivery vehicles can also be added to paper products forthe topical application of cargo moieties to skin. Examples of paperproducts that can include geodate delivery vehicles of the inventioninclude baby care products, i.e, diapers or baby wipes, tissues, toiletpaper, antibacterial or antiperspirant towelettes, napkins, papertowels, bandaids, gauze pads, and feminine hygiene products.

[0100] An artisan can determine without undue experimentation theoptimal lipid to cargo moiety and/or hydrophobic domain ratios for aspecific purpose. Formation of geodate delivery vehicles is monitoredreadily. Then, the preparation can be administered to the targeted hostto ascertain the nature and tenor of the biologic response to theadministered composition. It should be evident that the optimized ratiofor any one use may range from a high ratio, for example, to minimizethe use of a rare cargo moiety, to a low ratio to obtain maximal amountof cargo moiety in the vehicle. Because the vehicle can accept a largeload of cargo moiety, the amount of cargo moiety can vary greatlydepending on need.

[0101] The present invention also provides a method of manufacturing ageodate delivery vehicle for a cargo moiety. The method generallyincludes the step of: mixing a lipid, an aqueous solution and ahydrophobic material, such that a geodate delivery vehicle is formed,which includes a lipid monolayer disposed about a hydrophobic domain.

[0102] An alternate method of forming a geodate delivery vehicleincludes mixing a lipid and a hydrophobic material, e.g., by kneading,such that one or more geodate delivery vehicles are formed. This methodcan be advantageous, for example, when an aqueous environment is notdesired. Fragile cargo moieties are often sensitive to moisture, whichcan cause decomposition upon prolonged exposure. A non-aqueous methodfor forming geodate delivery vehicles, therefore, would be desirable

[0103] At a low lipid to hydrophobic domain ratio, the lipids tend toform micelles in the water; at a higher concentration the lipids willform lipid monolayers about the hydrophobic globule. Preferably, thelipid to hydrophobic domain ratio is between 5:1 and about 10:1.

[0104] The method can further include the step of adding a cargo moiety,wherein the cargo moiety associates with the hydrophobic domain. Theagent can be added prior to or after emulsifying the mixture to form thelipid monolayer about the hydrophobic domain. Alternatively, thehydrophobic domain may itself be a cargo moiety, e.g., fish oil.

[0105] The methods of the invention can include the step of adding acation to the emulsion to form a lipid strata about a geodate deliveryvehicle. The lipid strata can be maintained in the emulsion. Optionally,the method can include the step of extracting the precipitate from theemulsion to form a solid or semi-solid, e.g., a powder. The geodatedelivery vehicle can be harvested from the suspension by filtration,centrifugation or other techniques, and dried to a powder. As shownbelow, the geodates can be extracted from suspension using commercial,large-scale or large batch equipment, e.g., spray dryers or fluid beddryers. Geodes recovered with such equipment can experience extremetemperatures, e.g., 400° F., for prolonged periods of time withoutdegrading the geode structure or its cargo.

[0106] Alternatively, the geodates can be dried using an apparatus whichuses high pressure and hot air to form a powder. The high pressurecreates a mist from the geodate suspension, which enters a chamber fromthe top. The hot air enters the camber from the bottom and blows seedcrystals into the center of the chamber. When the mist and the seedcrystals meet, geodates coat the seed crystal, and powder forms.

[0107] Use of the delivery vehicles of the present invention, e.g.,geodes, can result in an increase in the amount of active ingredientdelivered versus that which can be achieved with conventional food ordrug preparations. For example, the delivery vehicles of the presentinvention can result in a 20%, 40%, 50%, 60%, 100%, 200% . . . 1000% . .. 10,000% increase in the active (undegraded) ingredient deliveredversus use of the cargo directly in the preparation of the drug, food,beverage, etc.

[0108]FIG. 1 illustrates an exemplary method of manufacturing a geodatedelivery vehicle in accordance with the present invention. The lipid(e.g., a phospholipid) is represented in liposomes as open rings, andindividually and in lipid monolayer arrangements as hairpin-likestructures indicating the hydrophilic head and hydrophobic tail portionsof a typical phospholipid. The hydrophobic domains (e.g., oil droplets),are represented by shaded circles. Lipid strata is represented withhatching. The phospholipid and oil droplets are emulsified to create astable emulsion that includes liposomes as well as geodate deliveryvehicles that each include a lipid monolayer disposed about a droplet ofoil. A cation is added (e.g., calcium), and the liposomes collapse intoa strata of alternating liposome bilayer and calcium layers. In FIG. 1,several crystals are depicted, each one capturing several geodatedelivery vehicles. Optionally, the crystals can be removed from thesuspension (not shown).

[0109] To further illustrate the present invention, FIGS. 2A-D includefour images of DOPS and olive oil interacting in an aqueous buffer.FIGS. 3A and B are two images of geodate delivery vehicles including aDOPS monolayer disposed about Amphotericin B in olive oil. FIG. 4 is animage of a DOPS geodate delivery vehicle that includes a DOPS monolayerabout fluorescent Amphotericin B interacting with olive oil. FIGS. 5 and6 are images of a lipid strata encrusting olive oil, where the lipid isRhodamine-labeled DOPS. FIGS. 7 and 8 are images of a geodate deliveryvehicle that includes a fluorescent DOPS monolayer disposed about oliveoil, after a lipid strata formed with calcium was removed with achelating agent.

[0110] The natural composition of the preparations of the presentinvention reduces the risks associated with other delivery methods suchas methods using unnatural chemicals or methods using infectious viralvector systems. The preparations are manufactured relatively easily andinexpensively, and are compatible with a wide range of cargo moieties.The preparations can be delivered orally in a suspension or vehicle suchas a food vehicle (e.g., liquid or solid food items). Thus, thepreparations of the present invention may eliminate the need for painfuland difficult injections. Moreover, the preparations are not restrictedto prescription drugs, but may also be used to deliver over-the-countermedication or other agents, such as vitamins, minerals or othernutrients.

[0111] An example of such cargo moieties are omega-3 fatty acids, whichare found mainly in fish oils and other fish products. Omega-3 fattyacids have been implicated in increased disease resistance and fertilityin animals, and they are shown to have a significantly positive effecton cholesterol and overall cardiovascular health in human beings. See,for example, Daviglus et al. N Engl J Med. 336: 1046-1053 (1997). One ofthe complications of incorporating them directly into food, however, istheir noticeable odor and taste.

[0112] The present invention provides a means for masking flavors andodors, such as those associated with omega-3 fatty acids, byencapsulation within a lipid strata. For example, omega-3 fattyacid-geodes have been added to beverages such as soy milk, milk, liquidyogurt, grapefruit juice, orange juice, smoothies, sports drinks, softdrinks, tea, coffee, and iced coffee. In each case, there was noalteration in taste or odor of the beverage, i.e., the characteristicfish odor of the omega-3 fatty acid was not discernable. Similarly,omega-3 fatty acid geodes can be used in goods that are then baked orcooked, such as cakes, muffins, pasta noodles, soups and cookies withoutalteration in taste or odor.

[0113] The present invention is also particularly advantageous for thedelivery of unstable cargo moieties such as beta-carotene. Beta-caroteneacts as an antioxidant by quenching singlet oxygen and other freeradicals. Unfortunately beta-carotene and other carotenoids are highlysusceptible to oxidation prior to incorporation into the body. Thisphenomenon is observed as a bleaching of the deep orange color. Britton,FASEB J. 9: 1551-1558 (1995).

[0114] The present invention provides beta-carotene with an oxygen-freeenvironment for storage before use. Surprisingly, the beta-carotenemaintained activity even after exposure to extreme temperatures andpressures which normally would degrade it. FIG. 18 is an the image ofbeta-carotene geodes contained within muffins baked at approximately350° F. for about twenty minutes. The activity is indicated by thered-orange color observed in the muffins. Additionally, beta-carotenegeodes can be incorporated into other baked or cooked items andbeverages.

[0115] In another embodiment, the geodate delivery vehicles can beemployed to deliver nonsteroidal anti-inflammatory drugs (NSAIDS),typically used to treat inflammation, muscle strains, and high fever.NSAIDS function by inhibiting cyclooxygenase-1 (COX1) andcyclooxygenase-2 (COX2). COX1 enzymes are responsible for protecting thelining of the stomach and COX2 enzymes are responsible for theproduction of prostaglandins, which are important in the inflammatoryprocess. Unfortunately, commercially available preparations of NSAIDSare active against both COX1 and COX2, and therefore have unwanted sideeffects such as ulcers, upset stomach or nausea.

[0116] Ibuprofen and naproxen are two of the more widely used and wellknown NSAIDS commonly used to relieve pain and fever. Low doses ofibuprofen are used to control pain, but inflammation can not beregulated without a higher dosage, which often causes stomach upset,diarrhea, dizziness, drowsiness, gas, heartburn, or headache, andoccasionally more serious side effects such as kidney toxicity orjaundice. Naproxen is used to treat both pain, and inflammation;however, diarrhea, constipation, dizziness, drowsiness, gas, heartburn,nausea, vomiting, headache, increased susceptibility to sunburn andringing in the ears are common side effects. A delivery vehicle,therefore, is needed to successfully deliver such NSAIDs to themacrophage without unwanted side effects.

[0117] Macrophages are important in the uptake of bacteria, fungi andparasites, and also play an important role in the inflammatory response.In addition to performing phagocytosis, macrophages have the potentialof being activated, a process that results in increased cell size,increased levels of lysosomal enzymes, more active metabolism, andgreater ability to phagocytose and kill ingested microbes. Afteractivation, macrophages secrete a wide variety of biologically activeproducts that, if unchecked, result in tissue injury and chronicinflammation. One of the secreted products, nitric oxide (NO), has comeinto the forefront as a mediator of inflammation.

[0118] Nitric oxide (NO) produced by inducible NOS plays an importantrole in inflammation, killing of bacterial pathogens, and tissue repair.NO formation increases during inflammation (i.e., in rheumatoidarthritis, ulcerative colitis, and Crohns disease), and several classicinflammatory symptoms, (i.e., erythema and vascular weakness) arereversed by NOS inhibitors. Nitric oxide has also been recognized asplaying a versatile role in the immune system. It is involved in thepathogenesis and control of infectious diseases, tumors, autoimmuneprocesses and chronic degenerative diseases.

[0119] The mechanism of action and side effects of NSAIDS, such asibuprofen and naproxen, are explained in part by the generation of NOfrom iNOS. Inhibition of iNOS expression and NO production by employingthe geodate delivery vehicles of the present invention could be a way totherapeutically decrease the inflammatory actions of these drugs.

[0120] Another aspect of the present invention is a method ofadministration of the preparations of the present invention.Accordingly, the present invention provides a method of treating asubject that can benefit from the administration of a cargo moiety,including the step of administering a geodate delivery vehiclecomprising a cargo moiety to a subject. The preparations of the presentinvention can be used to treat fungal infections (e.g., by delivery of aantifungal agent such as Amphotericin B), to treat or prevent HIVinfection (e.g., by delivery of a vaccine or a peptide to inducecellular immunity), to treat macular degeneration (e.g., bydelivery of anutriceutical), to treat inflammation (e.g., by delivery of ananti-inflammatory), to treat bacterial infections (e.g., by delivery ofantibiotics), and to provide nutrients (e.g., by delivery of vitamins,minerals or oils).

[0121] Accordingly, the present invention provides for both prophylacticand therapeutic methods of treating a subject at risk of (or susceptibleto) a disorder or having a disorder which can be treated with one ormore cargo moiety.

[0122] “Treatment”, or “treating” as used herein, is defined as theapplication or administration of a therapeutic agent (e.g., NSAIDS) to apatient, or application or administration of a therapeutic agent geodeof the invention to an isolated tissue or cell line, who has a diseaseor disorder, a symptom of disease or disorder or a predisposition towarda disease or disorder, with the purpose to cure, heal, alleviate,relieve, alter, remedy, ameliorate, improve or affect the disease ordisorder, the symptoms of the disease or disorder, or the predispositiontoward disease. “Treated,” as used herein, refers to the disease ordisorder being cured, healed, alleviated, relieved, altered, remedied,ameliorated improved or affected.

[0123] The methods of the present invention include methods ofadministering a cargo moiety to a subject or host, wherein the cargomoiety is associated with a geodate delivery vehicle of the invention.The geodate delivery vehicles of the present invention may beadministered orally, nasally, topically, intravenously, transdermally,buccally, sublingually, rectally, vaginally or parenterally.

[0124] The present invention provides a method for treating a subjectthat would behefit from administration of a composition of the presentinvention. Any therapeutic indication that would benefit from a cargomoiety, e.g., a drug or nutrient, can be treated by the methods of theinvention. Accordingly, the present invention provides methods oftreating a subject at risk for or having a disease or disorder which canbe treated with, for example, a protein, a small peptide, an antiviral,an anesthetic, an anti-infectious, an antifungal, an anticancer, animmunosuppressant, a steroidal anti-inflammatory, a non-steroidalanti-inflammatory, an antioxidant, an antidepressant which can besynthetic or naturally derived, a substance which supports or enhancesmental function or inhibits mental deterioration, an anticonvulsant, anHIV protease inhibitor, a non-nucleophilic reverse transcriptaseinhibitor, a cytokine, a tranquilizer and/or a vasodilatory agent. Themethod includes the step of administering to the subject a compositionof the invention, such that the disease or disorder is treated. Thedisease or disorder can be, e.g., inflammation, pain, infection, fungalinfection, bacterial infection, viral infection, parasitic disorders, animmune disorder, genetic disorders, degenerative disorders, cancer,proliferative disorders, obesity, depression, hair loss, impotence,hypertension, hypotension, dementia, senile dementia, or malnutrition.

[0125] The geodate delivery vehicles of the instant invention can beused to treat a variety of inflammations, including headache, arthritis,rheumatoid arthritis, osteoarthritis, acute gout-, acute or chronic softtissue damage associated with, e.g., a sports injury, tennis elbow,bursitis, tendonitis, acute or chronic back pain, such as a herniateddisc, carpal tunnel syndrome, glomerulonephritis, carditis, ulcerativecolitis, asthma, sepsis, and plantar fasciitis. The geodate deliveryvehicles of the invention can also be used to relieve pain resultingfrom surgery or other medical procedure. The geodate delivery vehiclesof the instant invention can further be used to treat a variety offungal infections, including candida, e.g., yeast infection, tinea,e.g., Athlete's foot, pityriasis, thrush, cryptococcal meningitis,histoplasmosis, and blastomycosis.

[0126] The geodate delivery vehicles of the instant invention can alsobe used to treat a variety of bacterial infections, including but notlimited to moderate to severe lower respiratory tract infections, skininfections, biliary tract infections, bone infections, antibioticprophylaxis, pseudomembraneous enterocolitis, central nervous systeminfections (e.g., meningitis and ventriculitis), intra-abdominalinfections (e.g., peritonitis), pneumonia, septicemia, soft tissueinfections, neutropaenic sepsis, joint infections, infectiveendocartidis, and urinary tract infections.

[0127] Exemplary bacteria that can be treated with the antibioticpreparation of the present invention include, but are not limited to,Staphylococcus aureus, Staphylococcus epidermidis, Streptococcuspyogenes, Streptococcus pneumoniae, Streptococcus Group D, Clostridiumperfringens, Haemophilus influenzae, Escherichia coli, Pseudomonasaeruginosa, and Klebsiella pneumoniae.

[0128] The above methods can be employed in the absence of othertreatment, or in combination with other treatments. Such treatments canbe started prior to, concurrent with, or after the administration of thecompositions of the instant invention. Accordingly, the methods of theinvention can further include the step of administering a secondtreatment, such as for example, a second treatment for the disease ordisorder or to ameliorate side effects of other treatments. Such secondtreatment can include, e.g., radiation, chemotherapy, transfusion,operations (e.g., excision to remove tumors), and gene therapy.Additionally or alternatively, further treatment can includeadministration of drugs to further treat the disease or to treat a sideeffect of the disease or other treatments (e.g., anti-nausea drugs).

[0129] With regard to both prophylactic and therapeutic methods oftreatment, such treatments may be specifically tailored or modified,based on knowledge obtained from the field of pharmacogenomics.“Pharmacogenomics”, as used herein, refers to the application ofgenomics technologies such as gene sequencing, statistical genetics, andgene expression analysis to drugs in clinical development and on themarket.

[0130] More specifically, the term refers the study of how a patient'sgenes determine his or her response to a drug (e.g., a patient's “drugresponse phenotype”, or “drug response genotype”). Thus, another aspectof the invention provides methods for tailoring an individual'sprophylactic or therapeutic treatment according to that individual'sdrug response genotype. Pharmacogenomics allows a clinician or physicianto target prophylactic or therapeutic treatments to patients who willmost benefit from the treatment and to avoid treatment of patients whowill experience toxic drug-related side effects.

[0131] The language “therapeutically effective amount” is that amountnecessary or sufficient to produce the desired physiologic response. Theeffective amount may vary depending on such factors as the size andweight of the subject, or the particular compound. The effective amountmay be determined through consideration of the toxicity and therapeuticefficacy of the compounds by standard pharmaceutical procedures in cellcultures or experimental animals, e.g., for determining the LD50 (thedose lethal to 50% of the population) and the ED50 (the dosetherapeutically effective in 50% of the population). The dose ratiobetween toxic and therapeutic effects is the therapeutic index and itmay be expressed as the ratio LD50/ED50. Compounds which exhibit largetherapeutic indices are preferred. While compounds that exhibit toxicside effects may be used, care should be taken to design a deliverysystem that targets such compounds to the site of affected tissue inorder to minimize potential damage to unaffected cells and, thereby,reduce side effects.

[0132] In yet another aspect, the invention provides kits or otherwisepackaged geodate delivery vehicles. In one embodiment, the inventionprovides a packaged geodate delivery vehicle including: a geodatedelivery vehicle of the invention packaged with instructions for addingthe vehicle to a food, beverage or personal care product. In anotherembodiment, the packaged geodate delivery vehicle is packaged withinstructions for incorporating a cargo moiety into the geodate deliveryvehicle.

[0133] This invention is further illustrated by the following exampleswhich should not be construed as limiting.

EXAMPLES

[0134] Materials and Methods

[0135] Imaging of Geodes

[0136] Phase contrast light microscopy and confocal microscopy (Olympus)were used to image suspensions, cochleates, and geodes, with and withoutthe aid of fluorescence, which can be used, e.g., in the future study ofcellular uptake and intracellular distribution of fluorescently labeledgeodes and cargo moieties. Confocal microscopy is particularlyadvantageous as it is a 3-dimensional digital imaging device that can beused to effectively view slices of cell culture.

[0137] Cell Lines and Culture Conditions

[0138] Mouse macrophage J774A.1 cell line was obtained from ATCC. Themacrophage cells were grown in monolayers in humidified air with 5% CO₂at 37° C. in 60 mm² Petri dishes (Corning) containing 5 mL of DMEMsupplemented with 10% FBS. For experiments, cells were harvested byscraping and were seeded into 96-well plates at a density of 5×10⁵cells.

Example 1 A Lipid Monolayer Preparation Of A Cargo Moiety

[0139] In a first vessel, a hydrophobic composition was prepared byvortexing dried Amphotericin B (fungal agent) and rhodamine (afluorescent marking agent) with olive oil until the amphotericin and therhodamine were integrally mixed with the olive oil. In a separatevessel, dried lipid was vigorously mixed in water to obtain a suspensionof liposomes in water. The hydrophobic composition was then added to twoportions of the liposome suspension in lipid-to-oil ratios of about 10:1and about 5:1, and vigorously mixed to form stable emulsions. Inspectionof both emulsions under a microscope revealed the formation of thehydrophobic composition encapsulated with a lipid monolayer andliposomes (FIGS. 2A-D, 3A-B and 4 depict similar emulsions). Theemulsions were stable and the hydrophobic domain did not coalesce. Sucha stable emulsion is illustrated in FIG. 1, wherein the stable emulsionincludes geodate delivery vehicles that include lipid monolayers formedabout the hydrophobic domains (dark shading), and liposomes.

Example 2 Lipid Monolayer Preparation Cargo Moiety Trapped in a LipidStrata

[0140] Calcium was added to the emulsions of Example 1. A crystallinestructure was observed to form about the lipid monolayer. Thecrystalline structure is believed to include the calcium and liposomes.Each crystal enveloped several encapsulated hydrophobic domain asdepicted schematically in FIG. 1. (FIGS. 5 and 6 depict similarstructures).

Example 3 Release Of Lipid Monolayer Preparation From Strata

[0141] EDTA was added to the emulsion of Example 2. The crystalstructure was observed to deteriorate such that the encapsulated domainremained, no longer encrusted by the crystalline structure. (FIGS. 7 and8 depict similar emulsions).

Example 4 Preparation of Beta-Carotene in Geodate Delivery Vehicle

[0142] 500 mg of soy phosphatidylserine (PS), 250 mg of 20%beta-carotene in soy oil, 10 mg alpha tocopherol (Vitamin E), and 240 mgof soy oil were weighed into a glass tube. A smooth emulsion wasprepared by vigorously mixing the sample at 45° C. Most of thebeta-carotene was observed to be incorporated into the oil droplets whenexamined by light microscopy. FIGS. 10A, 10B and 11 depict stablebeta-carotene, oil, and lipid emulsions.

[0143] 4.5 ml of sterile water was added to the glass tube with vigorousmixing. Microscopic examination revealed a stable emulsion with manydifferent size oil droplets with beta-carotene, a smaller amount of freeoil droplets, and many small moving particles.

[0144] 3.3 ml of 0.1 M calcium was added in droplets to the emulsionwith vigorous mixing. Examination of this preparation under themicroscope revealed vehicle comprising PS monolayers formed about oildroplets containing beta-carotene, captured or encrusted within a lipidstrata. FIG. 9 schematically illustrates the method used and the resultsobserved in this experiment.

Example 5 Beta-Carotene Geodate Delivery Vehicles with Casein

[0145] 12 g of soy phosphatidylserine (PS), 3 g of 20% beta-carotene insoy oil, and 0.2 g alpha tocopherol (Vitamin E), were weighed into aglass tube. A smooth emulsion was prepared by vigorously mixing thesample at room temperature while slowly adding 30-40 ml of sterilewater. Most of the beta-carotene was observed to be incorporated intothe oil droplets when examined by light microscopy.

[0146] 4.8 g casein was added to the emulsion, followed by an additional60-70 ml of sterile water (added dropwise). Microscopic examinationrevealed a stable emulsion with many different size oil droplets withbeta-carotene, a smaller amount of free oil droplets, and many smallmoving particles.

[0147] 60 ml of 0.5 M calcium chloride was added in droplets to theemulsion with vigorous mixing. Examination of this preparation under themicroscope revealed vehicle comprising PS monolayers formed about oildroplets containing beta-carotene, captured or encrusted within a lipidstrata. The final composition was calculated as follows

[0148] Final Composition:

[0149] 60% phosphatidylserine

[0150] 24% casein

[0151] 1% tocopherol

[0152] 3% beta-carotene

[0153] 12% soy oil

[0154] 100 ml sterile water

[0155] 60 ml 0.5M Ca⁺²

[0156]FIGS. 12A and 12B depict beta-carotene geodes before and aftermicronization. FIGS. 13A and 13B depict beta-carotene geodes insuspension. EDTA can be added to the suspension to release thebeta-carotene. FIGS. 13C and 13D depict the suspensions in 13A and 13B,respectively, after the addition of EDTA.

Example 6 Preparation of Fish Oil-Geodate Delivery Vehicles with Casein

[0157] 12 g of soy phosphatidylserine (PS), 3 g of 30% fish oil, oliveoil and 0.2 g alpha tocopherol (Vitamin E), were weighed into a glasstube. A smooth emulsion was prepared by vigorously mixing the sample at45° C. while slowly adding 30-40 ml of sterile water.

[0158] 4.8 g casein was added to the emulsion, followed by an additional60-70 ml of sterile water (added dropwise). Microscopic examinationrevealed a stable emulsion with many different size fish oil droplets.

[0159] 60 ml of 0.5 M calcium chloride was added in droplets to theemulsion with vigorous mixing. Examination of this preparation under themicroscope revealed vehicle comprising PS monolayers formed about thefish oil droplets, captured or encrusted within a lipid strata. Thefinal compositions were as follows.

[0160] Final Composition of Fish Oil-Geodes with Casein

[0161] 0.30% fish oil

[0162] 60% soy phosphatidylserine

[0163] 10% casein

[0164] 50 ml sterile water

[0165] 60 ml 0.1M Ca⁺²

[0166] Fish Oil/Casein Geodes with Olive Oil and Tocopherol

[0167] 30% fish oil (10% olive oil, 1% tocopherol)

[0168] 60% soy phosphatidylserine

[0169] 10% casein

[0170] 50 ml sterile water

[0171] 60 ml 0.1M Ca⁺²

[0172] Fish Oil/Casein Geodes with Olive Oil, Garlic, Curcumin andTocopherol

[0173] 30% fish oil (10% olive oil with garlic and curcumin, 1%tocopherol)

[0174] 60% soy phosphatidylserine

[0175] 10% casein

[0176] 50 ml sterile water

[0177] 60 ml 0.1MCa⁺²

Example 7 Preparation of Muffins Containing Beta-Carotene Geodes

[0178] Beta-carotene geodes were prepared as described in Example 4.These geodes were added to BETTY CROCKER SUPERMOIST white cake mix. Themix was cooked at 350° F. for 20 minutes. The muffins showed clusters ofbright orange geodes for at least 12 days subsequent to baking (see FIG.18).

Example 8 Preparation of Muffins Containing Fish Oil Geodes

[0179] Fish oil/olive oil/vitamin E geodes were prepared as described inExample 6. These geodes were added to BETTY CROCKER SUPERMOIST whitecake mix. The mix was cooked at 375° F. for about 20 minutes. Themuffins showed clusters of light green geodes, and had no noticeableodor for at least 24 hours after baking.

[0180] Geodes with only fish oil and without olive oil or vitamin E wereprepared as described in Example 6. The geodes were added to the whitecake mix and cooked in a microwave until brown (full power, about 45-50seconds). The muffins had no odor or adverse taste immediately aftercooking and after 24 hours.

Example 9 Preparation of Soy Milk Suspension Containing Fish Oil Geodes

[0181] Fish oil geodes were prepared as described in Example 6. Oneteaspoon of fish oil geodes were added to 3 oz. YOPLAIT soymilk. Uponshaking, the fish oil geodes remained in a suspended state. The soy milkexhibited no noticeable odor or taste.

Example 10 Topical Application of Beta-Carotene Geodes

[0182] Beta carotene geodes were prepared as described in Example 4,added to petrolatum and applied to the surface of the palm. The coatingwas resistant to water (see FIG. 19). Without wishing to be bound to anyparticular theory, it is believed that the geodes may have fused withthe stratum corneum of the epidermis.

Example 11 Preparation of Fish Oil Geodes Containing Casein and 5%Tocopherol Using CaCl₂ Powder

[0183] 10 g tocopherol (Vitamin E—Roche) and 50 g fish oil (RocheROPUFA) were placed in a large KITCHENAID blender and thoroughly mixedby stirring at low speed. 120 g of soy PS (Degussa) was then added tothe fish oil/V-E mixture followed by several small aliquots of sterilewater. 20 g of casein was then added into the container with fish/V-Eemulsion, and sterile water was slowly added with constant low speedstirring until a total of 2000 ml water was added. Microscopicexamination showed a stable emulsion with many different size oildroplets and many small moving particles. 35.5 g of calcium chloridepowder was added to the container while constantly stirring at lowspeed. The suspension was subsequently stirred for an additional 30minutes, after which the sample was transferred into a sterile amberbottle and stored as an emulsion until further use. A spray dryer and afluid bed dryer have been used to powderize the fish oil geodes. Finalcomposition: Component Weight % before Ca⁺ % after Ca⁺ fish oil    50 g25% 21.2% tocopherol    10 g  5%  4.2% soy PS   120 g 60% 50.9% casein   20 g 10%  8.5% CaCl 35.529 g 15.1% sterile water   2000 mL

Example 12 Preparation of Beta-Carotene Geodes Containing Casein and0.8% Tocopherol Using CaCl₂ Powder

[0184] 2.6 g tocopherol (Roche) and 39 g of 20% B-carotene in olive oil(Cognis) were placed in a large KITCHENAID blender and thoroughly mixedby stirring at low speed. 156 g of Soy PS (Degussa) was added into thecontainer with B-carotene/vitamin E followed by several small aliquotsof sterile water. A smooth emulsion was prepared by vigorously mixingthe sample. 62.4 g of casein was then added to the container with thebeta-carotene emulsion, followed by slow addition of sterile water untila total volume of 2080 ml was reached. Microscopic examination showed astable emulsion with many different size oil droplets containingbeta-carotene, some free oil droplets, and many small moving particles.57.3 g of calcium chloride powder was slowly added to the container andthe suspension was mixed thoroughly. The suspension was subsequentlystirred for an additional 30 minutes at low speed. A 2.0 ml aliquot ofthe final preparation was placed in a 50 ml sterile tube for HPLC assay.The assay indicated that greater than 90% of the beta carotene in thesample was contained within the geodes. The emulsion was thentransferred into a sterile amber bottle until further use.

[0185] In order to determine whether the geodes could be successfullydried to a powder (removed from suspension) using commercial large-scaleequipment and without compromising the active agent disposed therein,one batch was dried using fluid bed drying equipment by Glatt AirTechniques, Inc. (Ramsey, N.J.), and the other using spray dryingequipment by Spray-Tech (Ontario, Calif.). FIGS. 15, 16 and 17 aregraphs depicting the stability of beta-carotene geodes after fluid beddrying and spray drying. FIG. 15 shows the amount of beta-carotene inthe formulation of geodes in suspension, after fluid bed drying, andafter spray drying compared to a theoretical 100%. FIG. 17 shows similardata for beta-carotene geodes in suspension, after spray drying, and, acombination of two batched of spray-dried geodes. Concentrations of betacarotene after drying are slightly higher than expected based on theamount present in starting material, possibly due to some loss of othercomponents. Higher recoveries of beta carotene were observed in the oildroplets within the geodes as compared to beta carotene elsewhere. FIG.16 is a graph showing the concentration of various formulations ofgeodes stored for a 20 day period in the presence and absence of light.The bottom graph has been normalized to a starting concentration of 1,so that a comparison of formulations can be made. The decrease inconcentration in powder upon storage may be exaggerated due to thehygroscopic geode powder taking on water.

[0186] In the fluid bed process, the geodes experienced temperatures ofat least 85° C., and in the spray dried process, the geodes experiencedtemperatures of up to 375° F. to 400° F., in some instances (where thegeodes stuck to the processing equipment) for several hours. Thebeta-carotene in the geodes remained active for both batches, asindicated by the red-yellow color of the samples, including the geodesremoved from the processing equipment.

[0187] Images of the geodes indicated that the geodes were successfullyprepared and that the beta-carotene was still active (FIGS. 14A-D).FIGS. 14A and 14D are images of geodes extracted by spray drying, andFIGS. 14B and 14C are images of geodes extracted by fluid bed drying.

[0188] Samples of the geodes were subsequently exposed to light and airfor 2½ days, and no degradation of beta-carotene was observed. FinalComposition: Component Weight % before Ca⁺ % after Ca⁺ PS   156 g 60%49.2% casein  62.4 g 24% 19.7% tocopherol  2.6 g  1%  0.8% B-carotene  39 g  3%  2.5% (with olive oil) olive oil 12%  9.8% CaCl 57.33 g 18.1%sterile water  2080 ml

Example 13 Preparation of NSAID Geodes

[0189] NSAID (ibuprofen and/or naproxen) was thoroughly mixed in oliveoil (5% to 10% by weight of total geode mixture). Soy PS in a lipid todrug ratio of 10:1 was added to a test tube. The NSAID/olive oil mixturewas then added to the tube containing powdered soy PS, and a spatula wasused to thoroughly mix the powder with the oil.

[0190] Once a homogeneous paste formed, TES buffer (pH 7.4) was slowlyadded to the tube and vortexed for 10 to 15 minutes to further mix thesuspension. The sample was observed under an optical microscope toensure NSAID crystals were not free in suspension, but contained in theoil. Calcium chloride at 2:1 ratio to lipid was added to the stableemulsion. Sample was again observed under dark optical microscopy todetermine that geodes had formed and there were no free NSAID crystalsin the aqueous environment. The crystals were then stored under nitrogenat 4° C. until further use.

Prophetic Example Assay for Nitrite Concentration

[0191] Accumulated nitrite (NO₂ ⁻) in culture medium will be measuredusing an automated calorimetric assay based on the Griess reaction.Swierkosz, T. A., et al. Br. J. Pharmacol.; 114(7): 1335-42, 1995.Gross, S., et al. Biochem. Biophys. Res. Commun. 178, 823-829, 1991.Ryu, Y. S., et al. Biochem. Biophys. Res. Comm. 272, 758-764, 2000.J774A.1 mouse macrophages will be incubated with LPS (1 μg/ml) plusIFN-γ (10 μg/ml) in the presence or absence of free ibuprofen, ibuprofencochleates, ibuprofen geodes as prepared in Example 13, free naproxen,naproxen cochleates, naproxen geodes as prepared in Example 13, andempty cochleates for 15 hours. 100 μl of sample will be reacted with theGriess reagent at room temperature for 10 minutes. NO₂ ⁻ will then bedetermined by measuring the absorbance at 540 nm in a microplate reader.A standard curve will be obtained using known concentrations of sodiumnitrite. In all experiments, NO₂ ⁻ concentration in wells containingmedium only will also be measured as a blank control.

[0192] Equivalents

[0193] Those skilled in the art will recognize, or be able to ascertainusing no more than routine experimentation, many equivalents to thespecific embodiments of the invention described wherein. Suchequivalents are intended to be encompassed by the following, claims.

What is claimed is:
 1. A geodate delivery vehicle for a cargo moietycomprising: a lipid monolayer disposed about a hydrophobic domain; and alipid strata disposed about the lipid monolayer.
 2. A geodate deliveryvehicle for a cargo moiety comprising: a lipid monolayer disposed abouta hydrophobic domain, wherein the lipid monolayer comprises aphospholipid.
 3. The geodate delivery vehicle of claim 1 or claim 2,wherein the geodate delivery vehicle is suspended in an aqueousenvironment.
 4. The geodate delivery vehicle of claim 3, wherein thegeodate delivery vehicle is suspended in an emulsion.
 5. The geodatedelivery vehicle of claim 1 or claim 2, wherein the geodate deliveryvehicle is in powder form.
 6. The geodate delivery vehicle of any one ofthe preceding claims further comprising a cargo moiety associated withthe geodate delivery vehicle.
 7. The geodate delivery vehicle of claim6, wherein the cargo moiety is associated with the hydrophobic domain.8. The geodate delivery vehicle of claim 6, wherein the hydrophobicdomain is a cargo moiety.
 9. The geodate delivery vehicle of claim 6,comprising a cargo moiety associated with the lipid monolayer or thelipid strata.
 10. The geodate delivery vehicle of claim 6, wherein thehydrophobic domain comprises a cargo moiety associated with an oil orfat.
 11. The geodate delivery vehicle of any one of the preceding claimswherein the lipid comprises a negatively charged phospholipid.
 12. Thegeodate delivery vehicle of any one of the preceding claims wherein thelipid comprises at least about 50% negatively charged lipid.
 13. Thegeodate delivery vehicle of any one of the preceding claims wherein thelipid comprises at least about 75% negatively charged lipid.
 14. Thegeodate delivery vehicle of claim 6, wherein the cargo moiety is atleast one member selected from the group consisting of a vitamin, amineral, a nutrient, a micronutrient, an amino acid, a toxin, amicrobicide, a microbistat, a co-factor, an enzyme, a polypeptide, apolypeptide aggregate, a polynucleotide, a lipid, a carbohydrate, anucleotide, a starch, a pigment, a fatty acid, a monounsaturated fattyacid, a polyunsaturated fatty acid, a flavor substance, a flavoredessential oil or extract, a hormone, a cytokine, a virus, an organelle,a steroid or other multi-ring structure, a saccharide, a metal, ametabolic poison, an antigen, an imaging agent, a porphyrin, atetrapyrrolic pigment, and a drug.
 15. The geodate delivery vehicle ofclaim 14, wherein the drug is at least one member selected from thegroup consisting of a protein, a small peptide, a bioactivepolynucleotide, an antibiotic, an antiviral, an anesthetic, ananti-infectious, an antifungal, an anticancer, an immunosuppressant, asteroidal anti-inflammatory, a non-steroidal anti-inflammatory, anantioxidant, an antidepressant which can be synthetic or naturallyderived, a substance which supports or enhances mental function orinhibits mental deterioration, an anticonvulsant, an HIV proteaseinhibitor, a non-nucleophilic reverse transcriptase inhibitor, acytokine, a tranquilizer or a vasodilatory agent.
 16. The geodatedelivery vehicle of claim 14, wherein the drug is at least one memberselected from the group consisting of Amphotericin B, acyclovir,adriamycin, carbamazepine, ivermectin, melphalen, nifedipine,indomethacin, curcumin, ibuprofen, naproxen, estrogens, testosterones,steroids, phenyloin, ergotamines, cannabinoids, rapamycin, propanadid,propofol, alphadione, echinomycin, miconazole nitrate, teniposide,hexamethylmelamine, taxol, taxotere, 18-hydroxydeoxycorticosterone,prednisolone, dexamethazone, cortisone, hydrocortisone, piroxicam,diazepam, verapamil, vancomycin, tobramycin, geldanamycin,acetaminophen, aspirin, nystatin, rifampin, vitamin A acid, mesalamine,risedronate, nitrofurantoin, dantrolene, etidronate, caspofungin,nicotine, amitriptyline, clomipramine, citalopram, dothepin, doxepin,fluoxetine, imipramine, lofepramine, mirtazapine, nortriptyline,paroxetine, reboxitine, sertraline, trazodone, venlafaxine, dopamine,St. John's wort, phosphatidylserine, phosphatidic acid, amastatin,antipain, bestatin, benzamidine, chymostatin, 3,4-dichloroisocoumarin,elastatinal, leupeptin, pepstatin, 1,10-phenanthroline, phosphoramidon,ethosuximide, ethotoin, felbamate, fosphenyloin, lamotrigine,levitiracetam, mephenyloin, methsuximide, oxcarbazepine, phenobarbital,phensuximide, primidone, topirimate, trimethadione, zonisamide,saquinavir, ritonavir, indinavir, nelfinavir, or amprenavir.
 17. Thegeodate delivery vehicle of claim 14, wherein the polynucleotide is atleast one member selected from the group consisting of adeoxyribonucletic acid (DNA) molecule, a ribonucleic acid (RNA)molecule, an siRNA, a ribozyme, an antisense molecule, a morpholino, anda plasmid.
 18. The geodate delivery vehicle of claim 14, wherein thepolypeptide is at least one member selected from the group consisting ofcyclosporin, angiotensin I, II, or III, enkephalins and their analogs,ACTH, anti-inflammatory peptides I, II, or III, bradykinin, calcitonin,beta-endorphin, dinorphin, leucokinin, leutinizing hormone releasinghormone (LHRH), insulin, neurokinins, somatostatin, substance P, thyroidreleasing hormone (TRH), and vasopressin.
 19. The geodate deliveryvehicle of claim 14, wherein the antigen is at least one member selectedfrom the group consisting of a carbohydrate, envelope glycoproteins fromviruses, an animal cell membrane protein, a plant cell membrane protein,a bacterial membrane protein and a parasitic membrane protein.
 20. Thegeodate delivery vehicle of claim 14, wherein the nutrient is at leastone member selected from the group consisting of micronutrients,vitamins, minerals, fatty acids, polyunsaturated fatty acids, aminoacids, fish oils, fish oil extracts, resveratrol, biotin, choline,inositol, ginko, and saccharides.
 21. The geodate delivery vehicle ofclaim 20, wherein the nutrient is a phytochemical or a zoochemical. 22.The geodate delivery vehicle of claim 14, wherein the nutrient is atleast one member selected from the group consisting of beta-carotene,lutein, zeaxanthine, quercetin, silibinin, perillyl alcohol, genistein,sulfurophane, lycopene, omega-3 and omega-6 fatty acids.
 23. The geodatedelivery vehicle of claim 14, wherein the vitamin is at least-one memberselected from the group consisting of vitamins A, B, B1, B2, B3, B12,B6, B-complex, C, D, E, and K, vitamin precursors, carotenoids andbeta-carotene.
 24. The geodate delivery vehicle of claim 14, wherein themineral is at least one member selected from the group consisting ofboron, chromium, colloidal minerals, colloidal silver, copper,manganese, potassium, selenium, vanadium, vanadyl sulfate, calcium,magnesium, barium, iron and zinc.
 25. The geodate delivery vehicle ofclaim 14, wherein the fatty acid is at least one member selected fromthe group consisting of monounsaturated, polyunsaturated and saturatedfatty acids.
 26. The geodate delivery vehicle of claim 14, wherein thesaccharide or sweetener is at least one member selected from the groupconsisting of saccharine, isomalt, maltodextrine, aspartame, glucose,maltose, dextrose, fructose and sucrose.
 27. The geodate deliveryvehicle of claim 14, wherein the flavor substance is at least one oil orextract selected from the group consisting of oils and extracts ofcinnamon, vanilla, almond, peppermint, spearmint, chamomile, geranium,ginger, grapefruit, hyssop, jasmine, lavender, lemon, lemongrass,marjoram, lime, nutmeg, orange, rosemary, sage, rose, thyme, anise,basil, black pepper, tea or tea extracts, an herb, a citrus, a spice anda seed.
 28. The geodate delivery vehicle of any of the preceding claimswherein the geodate delivery vehicle further comprises an aggregationinhibitor.
 29. The geodate delivery vehicle of claim 28, wherein theaggregation inhibitor comprises casein or methylcellulose. 30 A packagedgeodate delivery vehicle comprising: a geodate delivery vehicle of anyone of the preceding claims packaged with instructions for adding thevehicle to a food, beverage or personal care product.
 31. A food itemcomprising the geodate delivery vehicle of any one of the precedingclaims.
 32. The food item of claim 31, wherein the food item is ananimal food item, a human food item, a nutrient bar, a snack food, abeverage, a domesticated animal food, a fish food, a poultry feed, a petfood, a dog food or a cat food.
 33. A personal care product comprisingthe geodate delivery vehicle of any one of the preceding claims.
 34. Thepersonal care product of claim 33, wherein the personal care product isa hair care product or a skin care product.
 35. A packaged geodatedelivery vehicle comprising: a geodate delivery vehicle of any one ofclaims 1-5 packaged with instructions for incorporating a cargo moiety.36. A pharmaceutical composition comprising a geodate delivery vehicleof any of one of claims 1-29 and a pharmaceutically acceptable carrier.37. A method of treating a subject that can benefit from theadministration of a cargo moiety, comprising the step of administering ageodate delivery vehicle comprising a cargo moiety to a subject.
 38. Themethod of claim 37, wherein the administration route is selected fromthe group consisting of mucosal, systemic, oral, intranasal,intraocular, intrarectal, intravaginal, intrapulmonary, intravenous,intramuscular, subcutaneous, transdermal and intradermal.
 39. The methodof claim 37, wherein the cargo moiety is administered to treatinflammation, pain, infection, fungal infection, bacterial infection,viral infection, parasitic disorders, an immune disorder, geneticdisorders, degenerative disorders, cancer, diabetes, insomnia,proliferative disorders, obesity, depression, hair loss, impotence,hypertension, hypotension, dementia, senile dementia, or malnutrition.40. The method of claim 37, wherein the subject can benefit fromadministration of a nutrient and the cargo moiety is a nutrient.
 41. Amethod of manufacturing a geodate delivery vehicle for a cargo moiety,the method comprising the step of: mixing a lipid, an aqueous solutionand a hydrophobic material, such that a geodate delivery vehicle isformed comprising a lipid monolayer disposed about a hydrophobic domain.42. The method of claim 41, comprising the step of mixing a cargomoiety, a lipid, an aqueous solution and a hydrophobic material, suchthat a geodate delivery vehicle is formed comprising a lipid monolayerdisposed about a hydrophobic domain associated with the cargo moiety.43. The method of claim 41 or claim 42, comprising the step of forming alipid strata about the lipid monolayer by adding a multivalent cation.44. The method of claim 43, wherein the multivalent cation comprisescalcium.
 45. The method of claim 44, comprising the step of drying thegeodate delivery vehicle to form a powder.
 46. The method of any one ofclaims 41-45, comprising associating the geodate delivery vehicle with apharmaceutically acceptable carrier.
 47. The method of any one of claims41-45, comprising adding the geodate delivery vehicle to a food item ora personal care product.
 48. A method of forming a geodate deliveryvehicle for a cargo moiety, the method comprising: mixing a lipidcomprising a phospholipid and a hydrophobic material such that a geodatedelivery vehicle is formed.